Caliper GX II System Application Evaluation - PerkinElmer

Novel Analytical Technology to Support
Biologics Development: Caliper GX II
System Application Evaluation
Dr. Mei Lin
Sr. Research Investigator II
Biologics Process and Product Development/
Analytical Development & Testing
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Outline
•Caliper Technologies GXII System Information
•Assessment of Molecules
Concentration and Purity Determination of
Monoclonal Antibody
Titer and Purity of Small Proteins (MW 10-
30KD)
Analysis of Pegylated Proteins
•Conclusion
•Acknowledgements
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Caliper Technologies GXII System
An automated lab-on
on-a-chip design that
utilizes microfluidic capillary electrophoresis
and 96/384 well plate layout in lieu of SDS-
PAGE, Western Blot, and DNA/RNA gels
Software generates virtual gel image based
on electropherogram.
Sizing Range: ~10 to 200kDa
Linear Dynamic Range: 5 to 2000ng/μL
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Monoclonal Antibody Stability Sample
(Non-Reduced)
Intact
mAb
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Monoclonal Antibody Stability Sample
(Reduced)
HC
LC
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Overlay of Monoclonal Antibody Stability
Sample (Reduced/ Non-Reduced)
HC
LC
Intact
mAb
System
Peak
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Protein Titer and Purity Evaluation
(MW ~ 12,000 Da)
(E.coli
Fermentation
Time Course Sample
T = 2 h
T = 22 h
Purified Protein
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Protein Titer and Purity Evaluation
(MW ~ 12,000 Da)
Protein
(R/NR)
Protein
at Various
Conc.
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Protein Titer and Purity Evaluation
(MW ~ 12,000 Da)
Peak Area
9000
8000
7000
6000
5000
4000
3000
2000
1000
0
Linearity
y = 4.2085x + 79.163
R 2 = 0.9928
0 500 1000 1500 2000 2500
Conc. (ug/mL)
• Conc.: 50 ug/mL
-
2 mg/mL
• Linear over three
orders of
magnitude
• Can function as
titer assay
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Pegylated Protein under Reduced and
Non-Reduced Condition
Purity and Lot to Lot Variability
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Conclusion
Caliper LabChip GXII System
High-throughput automated platform.
Rapid and accurate analysis of protein
samples for sizing, quantitation and purity.
Comparable resolution and sensitivity to
conventional CE-SDS.
Idea instrument for DOE experiments for
clone selection, protein expression,
purification and formulation development
condition optimization.
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Acknowledgements
Biologics Process and Product Development/
Analytical Development & Testing
• Joseph Calzada
Nara Dontha
• Ronna Scanlon Subbarao Mantha
• Barbara Twarowska Ying-Chen Chen
• Dr. Steve Hosselet
Dr. Mike Grace
Caliper Life Sciences
• Linda Gary Dr. Abbie Esterman
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